BFAT_VARPD
ID BFAT_VARPD Reviewed; 434 AA.
AC H8WR05;
DT 06-MAR-2013, integrated into UniProtKB/Swiss-Prot.
DT 16-MAY-2012, sequence version 1.
DT 03-AUG-2022, entry version 33.
DE RecName: Full=Beta-phenylalanine transaminase;
DE EC=2.6.1.-;
DE AltName: Full=Aromatic beta-amino acid aminotransferase;
DE AltName: Full=Beta-phenylalanine aminotransferase;
DE AltName: Full=VpAT;
OS Variovorax paradoxus.
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Comamonadaceae; Variovorax.
OX NCBI_TaxID=34073;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS)
RP IN COMPLEXES WITH PLP AND THE INHIBITOR 2-AMINOOXYACETATE, FUNCTION,
RP CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, ENANTIOSELECTIVITY, COFACTOR,
RP SUBUNIT, ACTIVITY REGULATION, BIOTECHNOLOGY, AND MUTAGENESIS OF ARG-41.
RC STRAIN=CBF3;
RX PubMed=23087034; DOI=10.1128/aem.02525-12;
RA Crismaru C.G., Wybenga G.G., Szymanski W., Wijma H.J., Wu B., Bartsch S.,
RA de Wildeman S., Poelarends G.J., Feringa B.L., Dijkstra B.W., Janssen D.B.;
RT "Biochemical properties and crystal structure of a beta-phenylalanine
RT aminotransferase from Variovorax paradoxus.";
RL Appl. Environ. Microbiol. 79:185-195(2013).
CC -!- FUNCTION: Aminotransferase that acts exclusively on beta-amino acids
CC and exhibits a broad substrate range in vitro, accepting meta-,
CC para- and, to a lesser extent, ortho-substituted beta-phenylalanine
CC derivatives as amino donors, and 2-oxoglutarate or pyruvate as amino
CC acceptors. Is highly enantioselective toward (S)-beta-phenylalanine (is
CC not active with (R)-beta-phenylalanine) and derivatives with different
CC substituents on the phenyl ring, allowing the kinetic resolution of
CC various racemic beta-amino acids to yield (R)-beta-amino acids with
CC >95% enantiomeric excess (ee). Highly prefers aromatic beta-amino acids
CC over aliphatic beta-amino acids; cannot use beta-alanine or beta-
CC glutamate as substrate. Is likely involved in the beta-phenylalanine
CC degradation pathway that allows V.paradoxus strain CBF3 to use beta-
CC phenylalanine as a sole nitrogen source. {ECO:0000269|PubMed:23087034}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-3-amino-3-phenylpropanoate + 2-oxoglutarate = 3-oxo-3-
CC phenylpropanoate + L-glutamate; Xref=Rhea:RHEA:47320,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:22731, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:68506; Evidence={ECO:0000269|PubMed:23087034};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-3-amino-3-phenylpropanoate + pyruvate = 3-oxo-3-
CC phenylpropanoate + L-alanine; Xref=Rhea:RHEA:47324,
CC ChEBI:CHEBI:15361, ChEBI:CHEBI:22731, ChEBI:CHEBI:57972,
CC ChEBI:CHEBI:68506; Evidence={ECO:0000269|PubMed:23087034};
CC -!- COFACTOR:
CC Name=pyridoxal 5'-phosphate; Xref=ChEBI:CHEBI:597326;
CC Evidence={ECO:0000269|PubMed:23087034};
CC Note=Binds 1 pyridoxal phosphate per subunit.
CC {ECO:0000269|PubMed:23087034};
CC -!- ACTIVITY REGULATION: Is inhibited by 2-aminooxyacetate (AOA), a mimic
CC of beta-alanine and a known inhibitor of aminotransferases.
CC {ECO:0000269|PubMed:23087034}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.5 mM for (S)-beta-phenylalanine;
CC KM=0.3 mM for 2-oxoglutarate;
CC Note=kcat is around 11 sec(-1) with (S)-beta-phenylalanine and 2-
CC oxoglutarate as substrates.;
CC pH dependence:
CC Has high activity over a broad pH range (4 to 11.2).;
CC Temperature dependence:
CC Optimum temperature is 55 degrees Celsius. Displays a 2-fold higher
CC activity at 55 degrees Celsius than at 30 degrees Celsius.;
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:23087034}.
CC -!- BIOTECHNOLOGY: Is an attractive template for future beta-
CC aminotransferase-engineering efforts toward the synthesis of
CC enantiomerically pure beta-amino acids. {ECO:0000269|PubMed:23087034}.
CC -!- SIMILARITY: Belongs to the class-III pyridoxal-phosphate-dependent
CC aminotransferase family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; HE608883; CCE46017.1; -; Genomic_DNA.
DR PDB; 4AO9; X-ray; 1.50 A; A/B=1-434.
DR PDB; 4AOA; X-ray; 2.28 A; A/B=1-434.
DR PDBsum; 4AO9; -.
DR PDBsum; 4AOA; -.
DR AlphaFoldDB; H8WR05; -.
DR SMR; H8WR05; -.
DR PRIDE; H8WR05; -.
DR BRENDA; 2.6.1.5; 234.
DR GO; GO:0030170; F:pyridoxal phosphate binding; IEA:InterPro.
DR GO; GO:0008483; F:transaminase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.40.640.10; -; 1.
DR Gene3D; 3.90.1150.10; -; 1.
DR InterPro; IPR005814; Aminotrans_3.
DR InterPro; IPR015424; PyrdxlP-dep_Trfase.
DR InterPro; IPR015421; PyrdxlP-dep_Trfase_major.
DR InterPro; IPR015422; PyrdxlP-dep_Trfase_small.
DR Pfam; PF00202; Aminotran_3; 1.
DR SUPFAM; SSF53383; SSF53383; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aminotransferase; Pyridoxal phosphate; Transferase.
FT CHAIN 1..434
FT /note="Beta-phenylalanine transaminase"
FT /id="PRO_0000421443"
FT BINDING 41
FT /ligand="(S)-3-amino-3-phenylpropanoate"
FT /ligand_id="ChEBI:CHEBI:68506"
FT /evidence="ECO:0000305"
FT BINDING 132..133
FT /ligand="pyridoxal 5'-phosphate"
FT /ligand_id="ChEBI:CHEBI:597326"
FT BINDING 300
FT /ligand="pyridoxal 5'-phosphate"
FT /ligand_id="ChEBI:CHEBI:597326"
FT MOD_RES 267
FT /note="N6-(pyridoxal phosphate)lysine"
FT MUTAGEN 41
FT /note="R->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:23087034"
FT HELIX 3..20
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 22..31
FT /evidence="ECO:0007829|PDB:4AO9"
FT TURN 32..34
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 36..38
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 42..44
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 52..57
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 60..63
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 68..73
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 74..77
FT /evidence="ECO:0007829|PDB:4AO9"
FT TURN 78..81
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 86..97
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 104..108
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 109..119
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 124..131
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 132..147
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 151..155
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 165..170
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 177..182
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 187..196
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 198..200
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 201..206
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 208..211
FT /evidence="ECO:0007829|PDB:4AO9"
FT TURN 212..214
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 215..217
FT /evidence="ECO:0007829|PDB:4AOA"
FT HELIX 220..233
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 236..240
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 244..247
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 252..256
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 261..266
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 267..270
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 276..280
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 282..285
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 286..288
FT /evidence="ECO:0007829|PDB:4AO9"
FT TURN 290..292
FT /evidence="ECO:0007829|PDB:4AO9"
FT TURN 300..303
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 305..317
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 321..345
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 350..354
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 357..362
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 370..373
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 378..390
FT /evidence="ECO:0007829|PDB:4AO9"
FT STRAND 400..402
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 409..425
FT /evidence="ECO:0007829|PDB:4AO9"
FT HELIX 427..429
FT /evidence="ECO:0007829|PDB:4AO9"
SQ SEQUENCE 434 AA; 46417 MW; C3F292AF6193C87E CRC64;
MTHAAIDQAL ADAYRRFTDA NPASQRQFEA QARYMPGANS RSVLFYAPFP LTIARGEGAA
LWDADGHRYA DFIAEYTAGV YGHSAPEIRD AVIEAMQGGI NLTGHNLLEG RLARLICERF
PQIEQLRFTN SGTEANLMAL TAALHFTGRR KIVVFSGGYH GGVLGFGARP SPTTVPFDFL
VLPYNDAQTA RAQIERHGPE IAVVLVEPMQ GASGCIPGQP DFLQALRESA TQVGALLVFD
EVMTSRLAPH GLANKLGIRS DLTTLGKYIG GGMSFGAFGG RADVMALFDP RTGPLAHSGT
FNNNVMTMAA GYAGLTKLFT PEAAGALAER GEALRARLNA LCANEGVAMQ FTGIGSLMNA
HFVQGDVRSS EDLAAVDGRL RQLLFFHLLN EDIYSSPRGF VVLSLPLTDA DIDRYVAAIG
SFIGGHGALL PRAN
